Human Enteroid Core

项目摘要-核心B 体外3-D“迷你肠道”培养系统，称为肠样细胞，由人类外科手术产生 标本或内窥镜活检，已被发现概括了多种细胞类型， 包括正常肠上皮的隐窝绒毛轴。这些技术的发展 肠样病变提供了一个令人兴奋的机会来推进我们对致病机理的理解 以及有益的微生物，其中许多可以预防或减轻 病原体引起的腹泻疾病，与肠道相互作用并引发反应 上皮组织。因此，人类肠道核心的远景目标是(1) 描述和比较从小肠的三个区域产生的人类肠样 (十二指肠、空肠、回肠)和结肠，从不同的患者身上建立它们的模型 人类腹泻疾病，(2)测试改善肠类疾病的新方法 正常肠上皮的模型，以及(3)提供这些肠样和特化生长 用于本申请中提出的其他项目(核心C、项目1、2和3)的试剂。 人类肠道核心的目标是提供一个集中化的设施来满足肠道 NAMSED提案涉及的所有调查人员的需求。的具体功能 核心将有两个功能：一个服务组件，它将(1)产生肠样培养 从我们现有的银行进行所有项目(2)从精选的新患者样本中建立肠样 (3)保持所有肠状突的冷冻库存(4)通道，并根据要求区分肠状突 (5)维持WNT、Nogin和r-Respondin产生细胞系，并标准化条件 肠样培养液(6)常规检测肠样细胞系的分化状态(7) 通过培训小组成员进行技术转让；开发部分，将(1) 开发和标准化生理测量作为肠样反应的指标(2)修改 增强或消除特定细胞类型或基因的存在(3)与 间充质细胞测试生长因子替代物并开发新结构 组件(例如，肠腔内翻，使得顶面朝外)(4) 加入免疫细胞以提高肠样病变模型的相关性。核心将是 对个别项目的需求作出反应，这些需求可能会随着研究项目的变化而变化 随着整个领域的发展和项目3中新平台的设计，继续进行。 将制定活动以满足我们和其他NASMED项目调查人员的需求。 我们的目标是在这些令人兴奋的努力中相互补充和合作，以发展肠样体 作为研究肠道疾病的模型。

Project Summary – Core B Ex-vivo 3-D “mini-intestine” culture systems, termed enteroids, generated from human surgical specimens or endoscopic biopsies, have been found to recapitulate the multiple cell types that comprise the crypt-villus axis of the normal intestinal epithelium. The development of these enteroids has provided an exciting opportunity to advance our understanding of how pathogenic as well as beneficial microorganisms, many of which can prevent or lessen the severity of pathogen-induced diarrheal illness, interact with and induce responses from the intestinal epithelium. Therefore, the long range objectives of the Human Enteroid Core are to (1) characterize and compare human enteroids generated from three regions of the small intestine (duodenum, jejunum, ileum) and colon, from different patients to establish their use as models of human diarrheal diseases, (2) test new approaches to improve the enteroids as a relevant model of normal intestinal epithelium, and (3) to provide these enteroids and specialized growth reagents for use in the other projects (Core C, Projects 1, 2, and 3) proposed in this application. The goal of the Human Enteroid Core is to provide a centralized facility to meet the enteroid needs of all of the investigators involved in the NAMSED proposal. The specific functions of the Core will have two functions: a Service component, which will (1) Produce enteroid cultures from our existing bank for all projects (2) Establish enteroids from select new patient samples (3) Maintain frozen stocks of all enteroids (4) Passage and differentiate enteroids per requests (5) Maintain WNT, noggin, and r-spondin producing cells lines and standardized conditioned media for enteroid culture (6) Routinely test the enteroid lines for differentiation status (7) Technology transfer through training group members; a Development component, which will (1) Develop and standardize physiological measures as indicators of enteroid responses (2) Modify enteroids to enhance or deplete the presence of specific cell types or genes (3) Co-culture with mesenchymal cells to test for substitution of growth factors and develop novel structural components (e.g., inversion of the enteroids such that the apical surface faces outward) (4) Incorporate immune cells to improve the relevance of the enteroid model. The Core will be responsive to needs of the individual Projects, which may change as the research projects proceed, as the overall field evolves and as new platforms are engineered in Project 3. New activities will be developed to meet the needs of our and other NASMED project investigators. Our goal is complementary and collaborative in these exciting efforts to develop the enteroids as models to study enteric disease.